Changes in the lamin A in situ proximity with LAP2α and emerin in response to R388P-LA expression.

<p><b>A,E)</b> Whole cell extracts of C2C12 cells either control (Ctrl) or overexpressing wild-type (WT) or R388P (RP) LA, tagged with GFP (left panel) or FLAG (right panel) were analysed by western blot using anti-GFP, anti-FLAG, anti-LAP2α, anti-emerin and anti-GAPDH antibodies, as indicated. <b>B-D, F-H)</b> C2C12 cells overexpressing WT (upper panel) or R388P (lower panels) GFP-LA or FLAG-LA were fixed, labelled with anti-GFP and anti-LAP2α or anti-FLAG and anti-emerin antibodies, and processed either for immunofluorescence <b>(B,F)</b> or proximity ligation assay (PLAssay) <b>(C,G)</b>, before observation at the confocal microscope. Scale bar, 10 μm. <b>D,H)</b> Quantification of PLA signals per nucleus among GFP/PLA or FLAG/LA positive cells as shown in <b>C,G)</b>. The graphs show the median intensity of the total PLA signals detected per nucleus (upper panel) and the median frequency of the PLA signals detected at the intranuclear periphery (lower panel). Boxes show first and third quartiles, bars are put according to Tukey method for n = 59 (WT) and 31 (R388P) nuclei for <b>D</b> and n = 67 (WT) and 74 (R388P) nuclei for <b>H</b> *** p < 0.001 (Mann Whitney test).